AA&MetaboliteQuantitationofMediaPostAALabeling
1) Remove 2 500 µl aliquots of supernatant into scintillation vials, add scintillation fluid and count.2) Aliquot 1.6 mls of the remaining supernatant into each of two pyrex glass tubes.3) Add 6 mls of chloroform-methanol, 1:2, and vortex vigorously.--> Still have a monophase and it may be best to allow this monophase to equilibrate for a little while.4) Remove all large debris by spinning at 3,000 rpm for 5 mins.......阅读全文
AA－－Metabolite－Quantitation－of－Media－PostAA－Labeling
1) Remove 2 500 µl aliquots of supernatant into scintillation vials, add scintillation fluid and count.2) Aliquot 1.6 mls of the remaining supernatant
AA－－Metabolite－Quantitation－of－Cell－Pellets－PostAA－Labeling
Extraction:1) Following spin, save supernatant for analysis. Be extremely careful not to disturb the pellet since it is somewhat dispersed.2) Immediat
Sphingomyelin－Quantitation－Postcholine－Labeling－of－HL60－Cells
Lipid Extraction1) Following the appropriate time of treatment, transfer 4.5 ml into each of two duplicate glass pyrex tubes and maintain on ice.2) Sp
Labeling－Tubulin－and－Quantifying－Labeling－Stoichiometry
Labeling Tubulin and Quantifying Labeling StoichiometryThis is a general procedure for coupling moieties with reactive succinimidyl esters to tubulin.
Raft－culture－media－(aka－Green’s－media)
Raft culture media (aka Green’s media)3 parts DMEM (including glutamine or glutamax)1 part Ham’s F125% FCSvarious supplements (not everybody uses the
Labeling－Tubulin－and－Quantifying－Labeling－Stoichiometry2
II. Labeling ProtocolThe procedure described below can be scaled down if desired. It is essential to perform all steps involving caged dyes under a sa
Yeast－Media
YEPD (non-selection)-1% yeast extract-2% peptone-1.5% agar (if needed for plates)After autoclaving add glucose to 2% by adding 100 ml of 20% solution
TUNEL－labeling
In Situ Cell Death (Apoptosis) Detection by TUNEL labelingby Boehringer Mannheim (Catalog No. 1684809), modified by Josiah N. Wilcox andJosé C. Rodrig
Biosynthetic－labeling
How long should cells be labeled? The ideal length of time to label cells depends on the protein of interest and the label that you are using. If you
Antibiotic－Concentration－in－Media
Antibiotics should be added to lower than 60 C broth, or filter sterilized: See note. "C'' refers to chromosomal resistance: "P" plasmid based
Tissue－Culture－Media
We use two different kinds of media. Most cells are grown in DMEM. A few lymphoid cell lines are grown in RPMI. Cells grown in DMEM must be grown in a
Detection－by－TUNEL－labeling
In Situ Cell Death (Apoptosis) Detection by TUNEL labelingby Boehringer Mannheim (Catalog No. 1684809), modified by Josiah N. Wilcox,José C. Rodriguez
CMFDA－Labeling－of－Platelet
OUTLINECMFDA (5-chloromethylfluorescein diacetate) is a lipophilic tracer that has an enormous advantage over ordinary tracers (e.g. FITC) because it
Immunofluorescence－Labeling－of－Cells
实验概要Antibodies are an important tool for demonstrating both the presence and the subcellular localization of an antigen. Cell staining is a very ver
BrdU－Labeling－Protocol
实验概要The thymidine analog, 5-bromo-2-deoxyuridine (BrdU),is a common reagent used for cell proliferation assays and for the detection of apoptotic
Arachidonic－Acid－Labeling
1) Grow cells to a density of 5-8 X 105 cells/ml in RPMI 1640 containing serum.2) Pellet cells and wash 1 time with room temperature PBS.3) Resuspend
Cell－Culture－Media－and－Solutions
Cell Culture Media and SolutionsDec. 18, 1990R. VeileAntimycotic/antibiotic media:To 1 liter of sterile RPMI 1640 with 2mM L-glutamine, add:165.0 ml f
Bacterial－Media－Solutions－and－Stocks
3 agar (200 ml)Add 6 grams agar to 200 ml deionized water. Autoclave to sterilize.1.6 agar (200 ml)Add 3.2 grams agar to 200 ml deionized water.Autocl
Yeast－Media,－Solutions－and－Stocks
Yeast Media:Note: Synthetic complete medium can be prepared by adding media supplements (see below).Medium using 6.7 g yeast nitrogen base without ami
Cell－Culture－Media－and－Solutions
Antimycotic/antibiotic media:To 1 liter of sterile RPMI 1640 with 2mM L-glutamine, add:165.0 ml fetal bovine serum, heat inactivated12.0 ml 200mM (100
MEDIA－FOR－EMBRYO－CULTURE－AND－MANIPULATION
M16 Medium (Protocol obtained from Karen Austen-Reed from SS Tan Laboratory, Anatomy Department)For oocyte maturation and routine culture of mouse emb
DNA－labeling－by－nick－translation
DNA labeling by nick translationreagents: DNA for labeling (concentration c > 150 ng/µl) modified nucleotides: Biotin-16-dUTP, Digoxigenin-11-dUTP, co
Multiple－studies－with－a－single－experiment:－The－Power－of－－...（一）
Multiple studies with a single experiment: The Power of Quantitative MultiplexingMultiple studies with a single experiment: The Power of Quantitative
寡核苷酸的相关操作
In this section, you will find techniques related to oligonucleotides, such as oligo purification by acrylamide gel, annealing two oligos to make doub
Embryonic－limb－bud－culture－in－media
Early in embryonic development, the region of the chick embryo which is determined to form a limb first differentiates from the rest of the embryo1. T
TISSUE－CULTURE－STOCK－SOLUTIONS－AND－MEDIA
MS MEDIUM FOR ARABIDOPSISTo 990 ml H2O add: Sucrose ........... 10.0 g MOPS .............. 0.5 g Agar .............. 8.0 g Adjust pH to 5.7
Retos－Buffers－－Media－Book
08-15 BufferCompoundMWFinal concentration200 mlGlycine75.07 0.1 M1.5 g1 M MgCl2 10 mM2 mlKOHto pH 9H2Oto 200 mlAAcetic Acid 1 MCompoundMWFinal concent
Basic－Method－for－Indirect－Immunofluorescence－Labeling
Basic Method for Indirect Immunofluorescence LabelingBackgroundThis is the method for indirect immunofluorescence labeling; that is, the antibodies do
Metabolic－Labeling－of－Cells－with－35S
1) Transfer to a 24 wells plate the desired colonies.2) Once the cells are attached (at least 8 hours after tripsinizing them) add ~1 ml ofDME (met-,
ThiolReactive－Probe－Labeling－Protocol
实验概要Invitrogen offers several fluorescent and biotinylated phalloidin and phallacidin derivatives for labeling F-actin. These phallotoxins, isolated